A Correlation between pH and Fluorescence Lifetime of 2′,7′-Bis(2-Carboxyethyl)-5(6)-carboxyfluorescein (BCECF) In Vivo and In Vitro

2007 ◽  
Vol 36 (2) ◽  
pp. 206-207 ◽  
Author(s):  
Takakazu Nakabayashi ◽  
Hui-Ping Wang ◽  
Kazuo Tsujimoto ◽  
Seiji Miyauchi ◽  
Naoki Kamo ◽  
...  
Keyword(s):  
Fluorescence Lifetime

In-vitro and in-vivo detection of p53 by fluorescence lifetime on a hybrid FITC-gold nanosensor

2010 ◽  
Author(s):  
L. Sironi ◽  
S. Freddi ◽  
L. D'Alfonso ◽  
M. Collini ◽  
T. Gorletta ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
In Vivo Detection

Two-dimensional fluorescence lifetime imaging for in-vitro and in-vivo application

1998 ◽  
Author(s):  
Paul M. W. French ◽  
Mark J. Dayel ◽  
Keith Dowling ◽  
Sam C. W. Hyde ◽  
M. J. Lever ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging ◽  
Two Dimensional ◽  
Lifetime Imaging

Analysis of energy metabolism of HeLa cancer cells in vitro and in vivo using fluorescence lifetime microscopy

2016 ◽  
Author(s):  
Maria Lukina ◽  
Marina Shirmanova ◽  
Varvara Dudenkova ◽  
Irina Druzhkova ◽  
Anastasia Shumilova ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Cancer Cells ◽  
Fluorescence Lifetime

scholarly journals In vivo fluorescence lifetime imaging captures metabolic changes in macrophages during wound responses in zebrafish

2020 ◽  
Author(s):  
Veronika Miskolci ◽  
Kelsey E Tweed ◽  
Michael R Lasarev ◽  
Emily C Britt ◽  
Courtney E McDougal ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Regulation ◽  
Immune Cell ◽  
Sterile Inflammation ◽  
Fluorescence Lifetime Imaging ◽  
Redox Ratio ◽  
Lifetime Imaging ◽  
Intracellular Metabolism

AbstractThe effector functions of macrophages across the spectrum of activation states in vitro are linked to profound metabolic rewiring. However, the metabolism of macrophages remains poorly characterized in vivo. To assess changes in the intracellular metabolism of macrophages in their native inflammatory microenvironment, we employed two-photon fluorescence lifetime imaging microscopy (FLIM) of metabolic coenzymes NAD(P)H and FAD. We found that pro-inflammatory activation of macrophages in vivo was associated with a decrease in the optical redox ratio [NAD(P)H/(NAD(P)H+FAD)] relative to a pro-resolving population during both infected and sterile inflammation. FLIM also resolved temporal changes in the optical redox ratio and lifetime variables of NAD(P)H in macrophages over the course of sterile inflammation. Collectively, we show that non-invasive and label-free imaging of autofluorescent metabolic coenzymes is sensitive to dynamic changes in macrophage activation in interstitial tissues. This imaging-based approach has broad applications in immunometabolism by probing in real time the temporal and spatial metabolic regulation of immune cell function in a live organism.SignificanceMetabolic regulation of macrophage effector functions has recently emerged as a key concept in immune cell biology. Studies rely on in vitro and ex vivo approaches to study macrophage metabolism, however the high plasticity of these cells suggest that removal from their native microenvironment may induce changes in their intracellular metabolism. Here, we show that fluorescence lifetime imaging microscopy of metabolic coenzymes captures dynamic changes in the metabolic activity of macrophages while maintaining them in their endogenous microenvironment. This approach also resolves variations on a single-cell level, in contrast to bulk measurements provided by traditional biochemical assays, making it a potentially valuable tool in the field of immunometabolism.

Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

2017 ◽  
Author(s):  
Alena Rudkouskaya ◽  
Nattawut Sinsuebphon ◽  
Xavier Intes ◽  
Joseph E. Mazurkiewicz ◽  
Margarida Barroso
Keyword(s):  
Tumor Cells ◽  
Fluorescence Lifetime ◽  
Receptor Ligand

scholarly journals In vitro and in vivo NIR Fluorescence Lifetime Imaging with a time-gated SPAD camera

2021 ◽  
Author(s):  
Jason T. Smith ◽  
Alena Rudkouskaya ◽  
Shan Gao ◽  
Juhi M. Gupta ◽  
Arin Ulku ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Near Infrared ◽  
Single Photon ◽  
In Vivo Studies ◽  
Fluorescence Lifetime Imaging ◽  
Iccd Camera ◽  
Lifetime Imaging ◽  
Nir Fluorescence

Near-infrared (NIR) fluorescence lifetime imaging (FLI) provides a unique contrast mechanism to monitor biological parameters and molecular events in vivo. Single-photon avalanche photodiode (SPAD) cameras have been recently demonstrated in FLI microscopy (FLIM) applications, but their suitability for in vivo macroscopic FLI (MFLI) in deep tissues remains to be demonstrated. Herein, we report in vivo NIR MFLI measurement with SwissSPAD2, a large time-gated SPAD camera. We first benchmark its performance in well-controlled in vitro experiments, ranging from monitoring environmental effects on fluorescence lifetime, to quantifying Förster Resonant Energy Transfer (FRET) between dyes. Next, we use it for in vivo studies of target-drug engagement in live and intact tumor xenografts using FRET. Information obtained with SwissSPAD2 was successfully compared to that obtained with a gated-ICCD camera, using two different approaches. Our results demonstrate that SPAD cameras offer a powerful technology for in vivo preclinical applications in the NIR window.

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